%0 Journal Article
%T Activity-based protein profiling of the hepatitis C virus replication in Huh-7 hepatoma cells using a non-directed active site probe
%A Ragunath Singaravelu
%A David R Blais
%A Craig S McKay
%A John Pezacki
%J Proteome Science
%D 2010
%I BioMed Central
%R 10.1186/1477-5956-8-5
%X The PS4¡Ô probe successfully targeted 19 active proteins in nine distinct protein families, some that were predominantly labeled in situ compared to the in vitro labeled cell homogenate. Nine proteins revealed altered activity levels during HCV replication. Some candidates identified, such as heat shock 70 kDa protein 8 (or HSP70 cognate), have been shown to influence viral release and abundance of cellular lipid droplets. Other differentially active PS4¡Ô targets, such as electron transfer flavoprotein alpha, protein disulfide isomerase A5, and nuclear distribution gene C homolog, constitute novel proteins that potentially mediate HCV propagation.These findings demonstrate the practicality and versatility of non-directed activity-based protein profiling (ABPP) to complement directed methods and accelerate the discovery of altered protein activities associated with pathological states such as HCV replication. Collectively, these results highlight the ability of in situ ABPP approaches to facilitate the identification of enzymes that are either predominantly or exclusively labeled in living cells. Several of these differentially active enzymes represent possible HCV-host interactions that could be targeted for diagnostic or therapeutic purposes.The hepatitis C virus (HCV) is the major causative agent of hepatitis that affects over three percent of the global population [1]. With no vaccines yet available and clinical treatments that have only limited success, many HCV-infected individuals develop chronic hepatitis which eventually progresses into liver steatosis, cirrhosis and hepatocellular carcinoma. HCV is the leading cause of liver disease and transplantation [2]. As the HCV genome is a single-stranded RNA molecule of only ~9.6 kb (Figure 1A) that encodes for ten mature viral proteins, HCV relies heavily on host factors for its propagation. During cell entry, HCV E1 and E2 viral proteins interact with four known host receptors CD81, claudin-1, SRB1, and occludin [3
%U http://www.proteomesci.com/content/8/1/5